SMAD4 promotes somatic-germline contact during murine oocyte growth

Author:

Granados-Aparici Sofia123ORCID,Yang Qin1,Clarke Hugh J14ORCID

Affiliation:

1. Research Institute, McGill University Health Centre

2. Present address: Cancer CIBER (CIBERONC)

3. Present address: Pathology Department, Medical School, University of Valencia-INCLIVA

4. Departments of Obstetrics and Gynecology and Biology, Division of Experimental Medicine, McGill University

Abstract

Development of the mammalian oocyte requires physical contact with the surrounding granulosa cells of the follicle, which provide it with essential nutrients and regulatory signals. This contact is achieved through specialized filopodia, termed transzonal projections (TZPs), that extend from the granulosa cells to the oocyte surface. Transforming growth factor (TGFβ) family ligands produced by the oocyte increase the number of TZPs, but how they do so is unknown. Using an inducible Cre recombinase strategy together with expression of green fluorescent protein to verify Cre activity in individual cells, we examined the effect of depleting the canonical TGFβ mediator, SMAD4, in mouse granulosa cells. We observed a 20–50% decrease in the total number of TZPs in SMAD4-depleted granulosa cell-oocyte complexes, and a 50% decrease in the number of newly generated TZPs when the granulosa cells were reaggregated with wild-type oocytes. Three-dimensional image analysis revealed that TZPs of SMAD4-depleted cells were longer than controls and more frequently oriented towards the oocyte. Strikingly, the transmembrane proteins, N-cadherin and Notch2, were reduced by 50% in SMAD4-depleted cells. SMAD4 may thus modulate a network of cell adhesion proteins that stabilize the attachment of TZPs to the oocyte, thereby amplifying signalling between the two cell types.

Funder

Canadian Institutes of Health Research

Natural Sciences and Engineering Research Council of Canada

Publisher

eLife Sciences Publications, Ltd

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